Hashiguchi Akiko, Yamaguchi Hisateru, Hitachi Keisuke, Watanabe Kazuo
Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan.
School of Nursing and Medical Care, Yokkaichi Nursing and Medical Care University, Yokkaichi 512-8045, Japan.
Plants (Basel). 2021 Jan 8;10(1):115. doi: 10.3390/plants10010115.
spp. is an economically important vegetable crop with high stress-tolerance and health benefits. However, proteomic analysis of the plant has been difficult due to the composition of its succulent cladodes; the abundant polysaccharides interfere with protein extraction. To facilitate proteomic analysis of this plant, we present a rapid and simple protein extraction method for (L.) Miller. The optimized method produced highly reproducible protein patterns and was compatible with a gel-free quantitative workflow without the need for additional purification. We successfully analyzed the cladode mesocarp and exocarp tissues, resulting in the identification of 319 proteins. In addition, we used this method to examine the relative changes in the proteome in response to salt stress to determine whether physiological changes could be captured. Qualified observations were obtained, revealing that salt stress increased phosphoenolpyruvate carboxylase abundance and decreased ribulose-bisphosphate carboxylase in young plants. These findings suggest that Crassulacean acid metabolism is promoted under salinity. This study highlights the efficacy of our optimized protein extraction method for elucidating the metabolic adaptations of using gel-free proteomic analysis.
景天属植物是一种具有重要经济价值的蔬菜作物，具有高抗逆性且有益健康。然而，由于其肉质叶状枝的组成，对该植物进行蛋白质组学分析一直很困难；丰富的多糖会干扰蛋白质提取。为便于对这种植物进行蛋白质组学分析，我们提出了一种针对景天属（L.）米勒植物的快速简便的蛋白质提取方法。优化后的方法产生了高度可重复的蛋白质图谱，并且与无需额外纯化的无凝胶定量工作流程兼容。我们成功分析了叶状枝的中果皮和外果皮组织，鉴定出319种蛋白质。此外，我们使用该方法研究景天属蛋白质组对盐胁迫的相对变化，以确定是否能捕捉到生理变化。获得了有价值的观察结果，表明盐胁迫会增加幼嫩景天属植物中磷酸烯醇式丙酮酸羧化酶的丰度，并降低核酮糖-1,5-二磷酸羧化酶的含量。这些发现表明，盐度下景天酸代谢得到促进。本研究强调了我们优化的蛋白质提取方法在通过无凝胶蛋白质组学分析阐明景天属植物代谢适应性方面的有效性。
